Semi-quantitative RT-PCR analysis of LIM mineralization protein 1 and its associated molecules in cultured human
Qi Zhang1, Xiaoying Wang, Zhuo Chen
1Key Laboratory for Oral Biomedical Engineering of Ministry of Education and Department of Cariology & Endodontics, Luoyu Road 237, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei Province 430079, PR China.
Objective:
LIM mineralization protein 1 (LMP-1), an intracellular signaling molecule, regulates osteoblast differentiation and maturation, as well as bone formation. However, the role of LMP-1 in the differentiation of human dental pulp cells and formation of dentin has not been determined. The study was to investigate the expression of LMP-1, the related proteins, such as bone morphogenetic proteins 2, 6 and 7 (BMP-2, BMP-6 and BMP-7), and core binding factor alpha 1 (Cbfa1) during the differentiation of cultured human dental pulp cells and the formation of mineralized nodules.
Design:
Differentiation of human dental pulp cells was induced by dexamethasone, asorbic acid and beta-glycerophosphate. The formation of mineralized nodules, was determined by Von Kossa staining and immunocytochemistry detection of dentin sialoprotein. Expression of LMP-1, the related proteins and the differentiation marker alkaline phosphatase (ALP) was analysed by reverse transcriptase-polymerase chain reaction (RT-PCR).
Results:
The expression of LMP-1, BMP-2, BMP-6, BMP-7 and Cbfa1 was significantly increased in the process of dental pulp cells differentiation and the formation of mineralized nodules, while the pattern of the expression was distinct.
Conclusions:
The elevated level of LMP-1, BMPs and Cbfa1 expression indicated they might play a role in the differentiation of human dental pulp cells and the formation of mineralized nodules.
